Casposase structure and the mechanistic link between DNA transposition and spacer acquisition by CRISPR-Cas.

Abstract:

:Key to CRISPR-Cas adaptive immunity is maintaining an ongoing record of invading nucleic acids, a process carried out by the Cas1-Cas2 complex that integrates short segments of foreign genetic material (spacers) into the CRISPR locus. It is hypothesized that Cas1 evolved from casposases, a novel class of transposases. We show here that the Methanosarcina mazei casposase can integrate varied forms of the casposon end in vitro, and recapitulates several properties of CRISPR-Cas integrases including site-specificity. The X-ray structure of the casposase bound to DNA representing the product of integration reveals a tetramer with target DNA bound snugly between two dimers in which single-stranded casposon end binding resembles that of spacer 3'-overhangs. The differences between transposase and CRISPR-Cas integrase are largely architectural, and it appears that evolutionary change involved changes in protein-protein interactions to favor Cas2 binding over tetramerization; this in turn led to preferred integration of single spacers over two transposon ends.

journal_name

Elife

journal_title

eLife

authors

Hickman AB,Kailasan S,Genzor P,Haase AD,Dyda F

doi

10.7554/eLife.50004

subject

Has Abstract

pub_date

2020-01-08 00:00:00

issn

2050-084X

pii

50004

journal_volume

9

pub_type

杂志文章

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